The largest database of trusted experimental protocols

Neo lite luciferase assay system

Manufactured by PerkinElmer
Sourced in Netherlands, United States

The Neo-Lite Luciferase Assay System is a luminescence-based detection kit designed for quantifying luciferase reporter gene expression. The system utilizes a proprietary luciferase substrate to generate a stable bioluminescent signal, allowing for sensitive and reliable measurement of luciferase activity in cell-based assays.

Automatically generated - may contain errors

5 protocols using neo lite luciferase assay system

1

Luciferase-Based Ad26 Neutralization Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Ad26 neutralizing antibody titers in serum were assessed using a luciferase-based VNA. Briefly, an E1/E3–deleted Ad26-luciferase reporter construct was added to 96-well half-area tissue culture–treated plates (Greiner) at a multiplicity of infection of 1,000, together with twofold serial dilutions of individual heat-inactivated serum samples starting at a 1:16 dilution in a total volume of 25 µl. After 1 h, A549 human lung carcinoma cells (catalog number ATCC CCL-185, obtained from the American Type Culture Collection) were added at a density of 1 × 104 cells/well. After incubation for 20 h at 37°C and 10% CO2, luciferase activity was measured using the Neo-Lite Luciferase Assay System (Perkin Elmer), and an EnVision multimode plate reader (Perkin Elmer). 90% neutralization titers (IC90) were defined as the maximum serum dilution that neutralized 90% of luciferase activity. Each serum sample was analyzed in duplicate.
+ Open protocol
+ Expand
2

Neutralizing Antibody Titers Measurement

Check if the same lab product or an alternative is used in the 5 most similar protocols
Ad26 and Ad35 NAb titers in serum were assessed using a luciferase-based VNA based on the method described in ref. 61 (link) A549 human lung carcinoma cells (catalog number ATCC CCL-185, obtained from the American Type Culture Collection, Manassas, VA) were plated at a density of 1 × 104 cells/well in 96-well black-and-white isoplates (Wallac, Turku, Finland). E1/E3-deleted Ad26- or Ad35-luciferase reporter constructs were then added at a multiplicity of infection (MOI) of 500, together with twofold serial dilutions of individual heat-inactivated cynomolgus macaque serum samples starting at a 1:64 dilution for Ad26, or a 1:32 dilution for Ad35 in 200 μl reaction volumes. After incubation for 24 h at 37 °C and 10% CO2, luciferase activity in cryolysed cells was measured using the Neo-Lite Luciferase Assay System (PerkinElmer, Groningen, Netherlands) on a BioTek Synergy Neo luminescence counter (BioTek). Ninety percent neutralization titers (IC90) were defined as the maximum serum dilution that neutralized 90% of luciferase activity. Each serum sample was analyzed in duplicate.
+ Open protocol
+ Expand
3

Quantifying Ad26 Neutralizing Antibodies

Check if the same lab product or an alternative is used in the 5 most similar protocols
Ad26 Nab titers in serum were assessed using a luciferase-based virus neutralization assay (VNA)50 (link). Briefly, heat-inactivated Cynomolgus Macaque serum samples was 2-fold serial diluted starting at a 1:32, 1:64 or 1:128 dilution for Ad26 (depending on the study, see figure legends for the exact start dilution). E1/E3 deleted Ad26-luciferase reported constructs were combined with serial diluted serum into a Tissue Culture treated Black and White Isoplate-96 (Perkin Elmer, Nederland B.V) at 500 to 1000 vp/cell. Plates were incubated for 30 minutes at RT, before A549 human lung carcinoma cells (ATCC® CCL-185™, American Type Culture Collection, Manassas, VA, USA) were added at a density of 1×104 cells/well. After incubation for 20 to 24 hours at 37 °C and 10% CO2, luciferase activity was measured using the Neo-Lite Luciferase Assay System (Perkin Elmer, Waltham, MA, USA) and a BioTek Synergy Neo luminescence counter (BioTek/Agilent, Santa Clara, CA, USA) or EnVision multimode plate reader (Perkin Elmer). A 90% neutralization titer (IC90) was defined as the maximum serum dilution that neutralized 90% of luciferase activity. Each serum sample was analyzed in duplicate.
+ Open protocol
+ Expand
4

NF-κB Activity Modulation by SDA and BAY

Check if the same lab product or an alternative is used in the 5 most similar protocols
To test the effect of SDA and BAY on NF-κB activity, androgen-sensitive LNCaP cells were transduced with lentiviral vectors carrying NF-κB response element sequence upstream of the luciferase gene sequence. Cells were treated with vehicle, SDA, or TNFα alone or with TNFα in combination with SDA or Bay 11-7280. Firefly luciferase activity was measured using the Neolite Luciferase Assay System (PerkinElmer, Downers Grove, IL).
+ Open protocol
+ Expand
5

Neutralizing Antibodies Against RSV

Check if the same lab product or an alternative is used in the 5 most similar protocols
VNA against RSV A2 was determined on serum of animals from study 2 using recombinant luciferase expressing RSV viruses9 (link). Five thousand A549 human lung carcinoma cells were added to each well of 96-well white half-area plates (Greiner Bio-One, Frickenhausen, Germany) containing 2.5 × 104 SFU/well of RSV-A2 viral particles encoding a luciferase reporter gene (resulting in a multiplicity of infection of 5), together with serial dilutions of individual heat-inactivated cynomolgus macaque serum. After incubation for 20 h at 37 °C and 10% CO2, luciferase activity in lysed cells was measured using the Neo-Lite Luciferase Assay System (Perkin Elmer) on a BioTek Synergy Neo luminescence counter (BioTek/Agilent). IC50 values were defined as the maximum serum dilution that neutralized luciferase activity by 50%. Each serum sample was analyzed in duplicate.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!